改良的大豆种皮过氧化物酶纯化方法及酶的底物氧化特性研究  

IMPROVED METHOD FOR PURIFICATION OF SOYBEAN HULL PEROXIDASE AND SUBSTRATE-OXIDATION OF THE ENZYME

作　　者：刘稳[1] 李杨[1] 段新源[1] 高培基[1] 卢雪梅[1] 

机构地区：[1]山东大学微生物技术国家重点实验室,山东济南250100

出　　处：《山东大学学报（理学版）》2001年第1期78-83,共6页Journal of Shandong University(Natural Science)

基　　金：国家自然科学基金! ( 2 990 60 0 5);国家重点实验室开放课题! (A1 4 99E0 6)

摘　　要：由大豆壳中分离出一种过氧化物酶 大豆种皮过氧化物酶 (soybeanhullperoxidase ,SHP ,EC 1 .1 1 .1 .7) ,经硫酸铵分级沉淀 ,DEAE SephadexA 50离子交换层析 ,Bio GelP 60凝胶过滤 ,DEAE SephadexA 2 5二次离子交换和SephadexG 1 0 0凝胶过滤纯化了该酶 .分子量为 3.8× 1 0 4u ,薄层等电聚焦电泳测得SHP等电点为 3.9.SHP的底物范围较宽 ,以H2 O2 为电子受体 ,可以氧化多种木素单体模型物 ,包括愈创木酚、2 ,3 二甲氧基酚、2 ,6 二甲氧基酚、3,4 二甲氧基酚、3,5 二甲氧基酚、1 ,2 ,4 三甲氧基苯、阿魏酸、咖啡酸、异丁子香酚和丁香醛联氮 ,提示该酶在木素的生物降解中有潜在的应用价值 .Soybean hull peroxidase(EC 1.11.1.7),isolated from Glycine max,was purified to electrophoretic homogeneity by combined consecutive treatments consisting of ammonium sulfate fractionation,column chromatography by gradient elutions from DEAE-Sephadex A-50,gel filtration on Bio-Gel P-60,rechromatography on DEAE-Sephadex A-25 and Sephadex G-100.The molecular weight of the enzyme,determined by SDS-PAGE,was 38000 and the isoelectric point was estimated to be around pH3.9 by IEF experiment.The enzyme possesses a wide substrate specificity along with a high potential and in the presence of hydrogen peroxide,can oxidize various lignin model compounds,including guaiacol,catechol,2,6-dimethoxyphenol,2,3-dimethoxyphenol,3,4-dimethoxyphenol,3,5-dimethoxyphenol,syringyl aldehyde,vanillylacetone,1,2,4-trimethoxybenzene,ferulic acid,isoeugenol,caffeic acid and syringaldazine,etc.This indicates that the enzyme has potential value in lignin degradation.

关 键 词：大豆种皮过氧化物酶 纯化 木素模型物 

分 类 号：Q741[生物学—分子生物学]